(1076-A) Liquid-Handler-Free Automation of Library Preparation for Decentralized Next Generation Sequencing

Abstract: Next generation sequencing (NGS) is increasingly accessible due to its decreasing costs and improved sequencing platforms. However, the library preparation for sequencing from gDNA remains a major bottleneck in the NGS workflow, particularly the PCR-free procedures. This creates a burden for small-to-medium scale laboratories to yield sequencing-ready samples. Hence, a stand-alone automated library preparation platform is presented here to further simplify the preparation of sequencing-ready samples. The capillary-based automation system requires minimal hands-on time of around 20-25 minutes of plate setup with single-point user interaction in the preparation process while the standard manual process requires ~3-4 hours with multiple interaction points. The PCR-free protocol run on the automated platform yielded 4.92 ± 1.832 ng/μL of ssDNA, comparable to manual library preparation of 5.12 ± 2.772 ng/μL . The sequence alignment of the two processes, automated and manual, had negligible differences to the hg38 reference genome at an average of 98.30% and 98.38% alignments, respectively. The automation platform has the potential to decentralize the NGS workflow making it more accessible to a wider range of users, particularly to researchers in local clinical laboratories, developing countries, and remote locations, where access to traditional NGS facilities is limited.